MBio Diagnostics, Inc., in collaboration with the University of California, San Diego, and PATH (Seattle, WA), proposes to develop a low-cost, no amplification, nucleic acid diagnostic technology for respiratory pathogens. The proposed system will address two major public health needs. First, it will establish a readily configurable diagnostic tool that can be rapidly deployed for public health screening and diagnostic efforts during a bioterror event or new pandemic emergencies. Second, the program will yield a viral RNA based flu rapid test that will offer significant sensitivity and specificity improvements over commercially available, antigen-based rapid diagnostics tests. While PCR and mass spectrometry methods are sensitive and specific enough for new pathogen identification and disease diagnosis, costs and turnaround times associated with these technologies render them of only limited use in large-scale screening applications. Next generation diagnostic tools with laboratory quality performance, but that are rapid, robust, very low cost, and readily reconfigurable are needed. Here we propose a research partnership for the development of a simple, no amplification nucleic acid detection system for respiratory pathogens from the NIAID Priority Pathogen list. Research will focus on a novel integration of biological sample preparation and target detection within a single device. The core research emphasis is on an innovative assay approach that combines direct magnetic particle capture of target nucleic acid sequences and simultaneous labeling for immediate, high sensitivity fluorescence detection on a simple reader instrument. The model pathogen under this award will be influenza virus, and development will focus on multiplexed, viral RNA detection in clinical specimens, with a sample-to-answer turnaround time of less than 40 minutes. Research aims are to (1) Demonstrate multiplexed, multi-particle nucleic acid extraction and fluorescence detection in increasingly complex samples with 30,000 target copies/ml sensitivity;(2) Build a well characterized human clinical sample collection to be used for assay and system development;(3) Develop a robust, low cost rapid test system consisting of single use disposable cartridges and a simple reader device;(4) Demonstrate feasibility of the rapid test detection system in laboratory and field conditions, and establish clinical sensitivity and specificity performance relative to approved flu rapid tests;and (5) Extend the technology to other NIAID Priority pathogens. Because the proposed technology will have ongoing, FDA-approved uses, critical infrastructure and trained users will be in place in the event of a biodefense emergency. In addition to providing proof-of-concept results on this next generation diagnostic technology, the proposed research will generally advance the science of particle based assays and will establish a well characterized influenza clinical specimen repository of ongoing use to NIAID programs. PUBLIC HEALTH RELEVANCE: The proposed nucleic acid based diagnostic system will fill a critical unmet need for improved, low cost, accurate public health screening and diagnosis. The system will have an ongoing use for seasonal flu testing, and will be quickly converted into a dedicated diagnostic tool in the event of a bioterror attack or pandemic emergency.